Structural and functional dynamics of Escherichia coli ribonuclease II : initial studies using a novel fluorescence based system

Abstract:

Ribonuclease II (RNase II) is a bacterial enzyme responsible for 90% of the
exonucleolytic degradation of mRNA in bacteria, and has bacterial homologues known to
be involved in virulence. The goal of this project was to examine the structural dynamics
of RNase II using fluorescence. Prior to the beginning of this project, little was known
regarding the structural composition of RNase II – required information in the study of
structural dynamics. Consequently, the structure of RNase II was studied by constructing
a series of deletion mutants in order to map the domains. The publication of an atomic
resolution structure of RNase II allowed the project to move directly into the study of
RNase II structural dynamics as it degrades mRNA. As a step towards this, RNase II was
fluorescently labeled, and preliminary binding studies of DNA – a competitive inhibitor –
to RNase II using fluorescence were conducted.